The present invention relates to inhibitors of protein kinase, especially c-Jun N-terminal kinases (JNK), which are members of the mitogen-activated protein (MAP) kinase family. There are a number of different genes and isoforms which encode JNKS. Members of the JNK family regulate signal transduction in response to environmental stress and proinflammatory cytokines and have been implicated to have a role in mediating a number of different disorders. The invention also relates to methods f or producing these inhibitors. The invention also provides pharmaceutical compositions comprising the inhibitors of the invention and methods of utilizing those compositions in the treatment and prevention of various disorders.
Mammalian cells respond to extracellular stimuli by activating signaling cascades that are mediated by members of the mitogen-activated protein (MAP) kinase family, which include the extracellular signal regulated kinases (ERKs), the p38 MAP kinases and the c-Jun N-terminal kinases (JNKs). MAP kinases (MAPKs) are activated by a variety of signals including growth factors, cytokines, UV radiation, and stress-inducing agents. MAPKs are serine/threonine kinases and their activation occur by dual phosphorylation of threonine and tyrosine at the Thr-X-Tyr segment in the activation loop. MAPKs phosphorylate various substrates including transcription factors, which in turn regulate the expression of specific sets of genes and thus mediate a specific response to the stimulus.
One particularly interesting kinase family are the c-Jun NH2-terminal protein kinases, also known as JNKs. Three distinct genes, JNK1, JNK2, JNK3 have been identified and at least ten different splicing isoforms of JNKs exist in mammalian cells [Gupta et al., EMBO J., 15:2760-70 (1996)]. Members of the JNK family are activated by proinflammatory cytokines, such as tumor necrosis factor-xcex1 (TNFxcex1) and interleukin-1 xcex2 (IL-1xcex2), as well as by environmental stress, including anisomycin, UV irradiation, hypoxia, and osmotic shock [Minden et al., Biochemica et Biophysica Acta, 1333:F85-F104 (1997)].
The down-stream substrates of JNKs include transcription factors c-Jun, ATF-2, Elk1, p53 and a cell death domain protein (DENN) [Zhang et al. Proc. Natl. Acad. Sci. USA, 95:2586-91 (1998)]. Each JNK isoform binds to these substrates with different affinities, suggesting a regulation of signaling pathways by substrate specificity of different JNKs in vivo (Gupta et al., supra).
JNKs, along with other MAPKs, have been implicated in having a role in mediating cellular response to cancer, thrombin-induced platelet aggregation, immunodeficiency disorders, autoimmune diseases, cell death, allergies, osteoporosis and heart disease. The therapeutic targets related to activation of the JNK pathway include chronic myelogenous leukemia (CML), rheumatoid arthritis, asthma, osteoarthritis, ischemia, cancer and neurodegenerative diseases.
Several reports have detailed the importance of JNK activation associated with liver disease or episodes of hepatic ischemia [Nat. Genet. 21:326-9 (1999); FEBS Lett. 420:201-4 (1997);J. Clin. Invest. 102:1942-50 (1998); Hepatology 28:1022-30 (1998)]. Therefore, inhibitors of JNK may be useful to treat various hepatic disorders.
A role for JNK in cardiovascular disease such as myocardial infarction or congestive heart failure has also been reported as it has been shown JNK mediates hypertrophic responses to various forms of cardiac stress [Circ. Res. 83:167-78 (1998); Circulation 97:1731-7 (1998); J. Biol. Chem. 272:28050-6 (1997); Circ. Res. 79:162-73 (1996);Circ. Res. 78:947-53 (1996); J. Clin. Invest. 97:508-14 (1996)].
It has been demonstrated that the JNK cascade also plays a role in T-cell activation, including activation of the IL-2 promoter. Thus, inhibitors of JNK may have therapeutic value in altering pathologic immune responses [J. Immunol. 162:3176-87 (1999); Eur. J. Immunol. 28:3867-77 (1998); J. Exp. Med. 186:941-53 (1997); Eur. J. Immunol. 26:989-94 (1996)].
A role for JNK activation in various cancers has also been established, suggesting the potential use of JNK inhibitors in cancer. For example, constitutively activated JNK is associated with HTLV-1 mediated tumorigenesis [Oncogene 13:135-42 (1996)]. JNK may play a role in Kaposi""s sarcoma (KS) because it is thought that the proliferative effects of bFGF and OSM on KS cells are mediated by their activation of the JNK signaling pathway [J. Clin. Invest. 99:1798-804 (1997)]. Other proliferative effects of other cytokines implicated in KS proliferation, such as vascular endothelial growth factor (VEGF), IL-6 and TNFxcex1, may also be mediated by JNK. In addition, regulation of the c-jun gene in p210 BCR-ABL transformed cells corresponds with activity of JNK, suggesting a role for JNK inhibitors in the treatment for chronic myelogenous leukemia (CML) [Blood 92:2450-60 (1998)].
JNK1 and JNK2 are widely expressed in a variety of tissues. In contrast, JNK3 is selectively expressed in the brain and to a lesser extent in the heart and testis [Gupta et al., supra; Mohit et al., Neuron 14:67-78 (1995); Martin et al., Brain Res. Mol. Brain Res. 35:47-57 (1996)]. JNK3 has been linked to neuronal apoptosis induced by kainic acid, indicating a role of JNK in the pathogenesis of glutamate neurotoxicity. In the adult human brain, JNK3 expression is localized to a subpopulation of pyramidal neurons in the CA1, CA4 and subiculum regions of the hippocampus and layers 3 and 5 of the neocortex [Mohit et al., supra]. The CA1 neurons of patients with acute hypoxia showed strong nuclear JNK3-immunoreactivity compared to minimal, diffuse cytoplasmic staining of the hippocampal neurons from brain tissues of normal patients [Zhang et al., supra]. Thus, JNK3 appears to be involved involved in hypoxic and ischemic damage of CA1 neurons in the hippocampus.
In addition, JNK3 co-localizes immunochemically with neurons vulnerable in Alzheimer""s disease [Mohit et al., supra]. Disruption of the JNK3 gene caused resistance of mice to the excitotoxic glutamate receptor agonist kainic acid, including the effects on seizure activity, AP-1 transcriptional activity and apoptosis of hippocampal neurons, indicating that the JNK3 signaling pathway is a critical component in the pathogenesis of glutamate neurotoxicity (Yang et al., Nature, 389:865-870 (1997)].
Based on these findings, JNK signalling, especially that of JNK3, has been implicated in the areas of apoptosis-driven neurodegenerative diseases such as Alzheimer""s Disease, Parkinson""s Disease, ALS (Amyotrophic Lateral Sclerosis), epilepsy and seizures, Huntington""s Disease, traumatic brain injuries, as well as ischemic and hemorrhaging stroke.
There is a high unmet medical need to develop JNK specific inhibitors that are useful in treating the various conditions associated with JNK activation, especially considering the currently available, relatively inadequate treatment options for the majority of these conditions.
Recently, we have described crystallizable complexes of JNK protein and adenosine monophosphate, including complexes comprising JNK3, in U.S. Provisional Application No. 60/084,056, filed May 4, 1998. Such information has been extremely useful in identifying and designing potential inhibitors of various members of the JNK family, which, in turn, have the described above therapeutic utility.
Much work has been done to identify and develop drugs that inhibit MAPKs, such as p38 inhibitors. See, e.g., WO 98/27098 and WO 95/31451. However, to our knowledge, no MAPK inhibitors have been shown to be specifically selective for JNKs versus other related MAPKs.
Accordingly, there is still a great need to develop potent inhibitors at JNKs, including JNK3 inhibitors, that are useful in treating various conditions associated with JNK activation.
It has now been found that compounds of this invention and pharmaceutical compositions thereof are effective as inhibitors of c-Jun N-terminal kinases (JNK). These compounds have the general formula I: 
where R1 is H, CONH2, T(n)xe2x80x94R, or T(n)xe2x80x94Ar2, n may be zero or one, and G, XYZ, and Q are as described below. Preferred compounds are those where the XYZ-containing ring is an isoxazole. Preferred G groups are optionally substituted phenyls and preferred Q are pyrimidine, pyridine or pyrazole rings.
These compounds and pharmaceutical compositions thereof are useful for treating or preventing a variety of disorders, such as heart disease, immunodeficiency disorders, inflammatory diseases, allergic diseases, autoimmune diseases, destructive bone disorders such as osteoporosis, proliferative disorders, infectious diseases and viral diseases. The compositions are also useful in methods for preventing cell death and hyperplasia and therefore may be used to treat or prevent reperfusion/ischemia in stroke, heart attacks, and organ hypoxia. The compositions are also useful in methods for preventing thrombin-induced platelet aggregation. The compositions are especially useful for disorders such as chronic myelogenous leukemia (CML), rheumatoid arthritis, asthma, osteoarthritis, ischemia, cancer, liver disease including hepatic ischemia, heart disease such as myocardial infarction and congestive heart failure, pathologic immune conditions involving T cell activation and neurodegenerative disorders.
This invention provides novel compounds, and pharmaceutically acceptable derivatives thereof, that are useful as JNK inhibitors. These compounds have the general formula I: 
wherein:
X-Y-Z is selected from one of the following: 
R1 is H, CONH2, T(n)xe2x80x94R, or T(n)xe2x80x94Ar2;
R is an aliphatic or substituted aliphatic group;
n is zero or one;
T is C(xe2x95x90O), CO2, CONH, S(O)2, S(O)2NH, COCH2 or CH2;
each R2 is independently selected from hydrogen, xe2x80x94R, xe2x80x94CH2OR, xe2x80x94CH2OH, xe2x80x94CHxe2x95x90O, xe2x80x94CH2SR, xe2x80x94CH2S(O)2R, xe2x80x94CH2(Cxe2x95x90O)R, xe2x80x94CH2CO2R, xe2x80x94CH2CO2H, xe2x80x94CH2CN, xe2x80x94CH2NHR, xe2x80x94CH2N(R)2, xe2x80x94CHxe2x95x90Nxe2x80x94OR, xe2x80x94CHxe2x95x90NNHR, xe2x80x94CHxe2x95x90NN(R)2, xe2x80x94CHxe2x95x90NNHCOR, xe2x80x94CHxe2x95x90NNHCO2R, xe2x80x94CHxe2x95x90NNHSO2R, -aryl, -substituted aryl, xe2x80x94CH2(aryl), xe2x80x94CH2(substituted aryl), xe2x80x94CH2NH2, xe2x80x94CH2NHCOR, xe2x80x94CH2NHCONHR, xe2x80x94CH2NHCON(R)2, xe2x80x94CH2NRCOR, xe2x80x94CH2NHCO2R, xe2x80x94CH2CONHR, xe2x80x94CH2CON(R)2, xe2x80x94CH2SO2NH2, xe2x80x94CH2(heterocyclyl), xe2x80x94CH2(substituted heterocyclyl), -(heterocyclyl), or -(substituted heterocyclyl);
each R3 is independently selected from hydrogen, R, COR, CO2R or S(O)2R;
G is R or Ar1;
Ar1 is aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclyl, or substituted heterocyclyl, wherein Ar1 is optionally fused to a partially unsaturated or fully unsaturated five to seven membered ring containing zero to three heteroatoms;
Qxe2x80x94NH is 
wherein the H of Qxe2x80x94NH is optionally replaced by R3; A is N or CR3;
U is CR3, O, S, or NR3;
Ar2 is aryl, substituted aryl, heterocyclyl or substituted heterocyclyl, wherein Ar2 is optionally fused to a partially unsaturated or fully unsaturated five to seven membered ring containing zero to three heteroatoms; and
wherein each substitutable carbon atom in Ar2, including the fused ring when present, is optionally and independently substituted by halo, R, OR, SR, OH, NO2, CN, NH2, NHR, N(R)2, NHCOR, NHCONHR, NHCON(R)21 NRCOR, NHCO2R, CO2R, CO2H, COR, CONHR, CON(R)2, S(O)2R, SONH2, S(O)R, SO2NHR, or NHS(O)2R, and wherein each saturated carbon in the fused ring is further optionally and independently substituted by xe2x95x90O, xe2x95x90S, xe2x95x90NNHR, xe2x95x90NNR2, xe2x95x90Nxe2x80x94OR, xe2x95x90NNHCOR, xe2x95x90NNHCO2R, xe2x95x90NNHSO2R, or xe2x95x90NR;
wherein each substitutable nitrogen atom in Ar2 is optionally substituted by R, COR, S(O)2R, or CO2R.
As used herein, the following definitions shall apply unless otherwise indicated. The term xe2x80x9caliphaticxe2x80x9d as used herein means straight chained, branched or cyclic C1-C12 hydrocarbons, preferably one to six carbons, which are completely saturated or which contain one or more units of unsaturation. For example, suitable aliphatic groups include substituted or unsubstituted linear, branched or cyclic alkyl, alkenyl, alkynyl groups and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl. The term xe2x80x9calkylxe2x80x9d and xe2x80x9calkoxyxe2x80x9d used alone or as part of a larger moiety refers to both straight and branched chains containing one to twelve carbon atoms. The terms xe2x80x9calkenylxe2x80x9d and xe2x80x9calkynylxe2x80x9d used alone or as part of a larger moiety shall include both straight and branched chains containing two to twelve carbon atoms. The terms xe2x80x9chaloalkylxe2x80x9d, xe2x80x9chaloalkenylxe2x80x9d and xe2x80x9chaloalkoxyxe2x80x9d means alkyl, alkenyl or alkoxy, as the case may be, substituted with one or more halogen atoms. The term xe2x80x9chalogenxe2x80x9d means F, Cl, Br, or I. The term xe2x80x9cheteroatomxe2x80x9d means N, O or S and shall include any oxidized form of nitrogen and sulfur, and the quaternized form of any basic nitrogen.
The term xe2x80x9carylxe2x80x9d, used alone or as part of a larger moiety as in xe2x80x9caralkylxe2x80x9d, refers to aromatic ring groups having five to fourteen members, such as phenyl, benzyl, 1-naphthyl, 2-naphthyl, 1-anthracyl and 2-anthracyl, and heterocyclic aromatic groups or heteroaryl groups such as 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxadiazolyl, 5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-pyrimidyl, 3-pyridazinyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 5-tetrazolyl, 2-triazolyl, 5-triazolyl, 2-thienyl, or 3-thienyl.
Aryl groups also include fused polycyclic aromatic ring systems in which a carbocyclic aromatic ring or heteroaryl ring is fused to one or more other rings. Examples include tetrahydronaphthyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolinyl, benzodiazepinyl, benzothiazolyl, benzooxazolyl, benzimidazolyl, isoquinolinyl, isoindolyl, acridinyl, benzoisoxazolyl, and the like. Also included within the scope of the term xe2x80x9carylxe2x80x9d, as it is used herein, is a group in which one or more carbocyclic aromatic rings and/or heteroaryl rings are fused to a cycloalkyl or non-aromatic heterocyclyl, for example, indanyl or tetrahydrobenzopyranyl.
The term xe2x80x9cheterocyclic ringxe2x80x9d or xe2x80x9cheterocyclylxe2x80x9d refers to a non-aromatic ring which includes one or more heteroatoms such as nitrogen, oxygen or sulfur in the ring. The ring can be five, six, seven or eight-membered and/or fused to another ring, such as a cycloalkyl or aromatic ring. Examples include 3-1H-benzimidazol-2-one, 3-1-alkyl-benzimidazol-2-one, 2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydropyranyl, 3-tetrahydropyranyl, 4-tetrahydropyranyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, 4-morpholino, 2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-piperazinyl, 2-piperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 4-thiazolidinyl, diazolonyl, N-substituted diazolonyl, 1-phthalimidinyl, benzoxane, benzotriazol-1-yl, benzopyrrolidine, benzopiperidine, benzoxolane, benzothiolane, and benzothiane.
A compound of this invention may contain a ring that is fused to a partially saturated or fully unsaturated five to seven membered ring containing zero to three heteroatoms. Such a fused ring may be an aromatic or non-aromatic monocyclic ring, examples of which include the aryl and heterocyclic rings described above.
An aryl group (carbocyclic and heterocyclic) or an aralkyl group, such as benzyl or phenethyl, may contain one or more substituents. Examples of suitable substituents on the unsaturated carbon atom of an aryl group include a halogen, xe2x80x94R, xe2x80x94OR, xe2x80x94OH, xe2x80x94SH, xe2x80x94SR, protected OH (such as acyloxy), phenyl (Ph), substituted Ph, xe2x80x94OPh, substituted xe2x80x94OPh, xe2x80x94NO2, xe2x80x94CN, xe2x80x94NH2, xe2x80x94NHR, xe2x80x94N(R)2, xe2x80x94NHCOR, xe2x80x94NHCONHR, xe2x80x94NHCON(R)2, xe2x80x94NRCOR, xe2x80x94NHCO2R, xe2x80x94CO2R, xe2x80x94CO2H, xe2x80x94COR, xe2x80x94CONHR, xe2x80x94CON(R)2, xe2x80x94S(O)2R, xe2x80x94SONH2, xe2x80x94S(O)R, xe2x80x94SO2NHR, or xe2x80x94NHS(O)2R, where R is an aliphatic group or a substituted aliphatic group.
An aliphatic group or a non-aromatic heterocyclic ring may contain one or more substituents. Examples of suitable substituents on the saturated carbon of an aliphatic group or of a non-aromatic heterocyclic ring include those listed above for the unsaturated carbon, such as in an aromatic ring, as well as the following: xe2x95x90O, =S, xe2x95x90NNHR, xe2x95x90NNR2, xe2x95x90Nxe2x80x94OR, xe2x95x90NNHCOR, xe2x95x90NNHCO2R, xe2x95x90NNHSO2R, or xe2x95x90NR.
A substitutable nitrogen on an aromatic or non-aromatic heterocyclic ring may be optionally substituted. Suitable substituents on the nitrogen include R, COR, S(O)2R, and CO2R, where R is an aliphatic group or a substituted aliphatic group.
Compounds derived by making isosteric or bioisosteric replacements of carboxylic acid or ester moieties of compounds described herein are within the scope of this invention. Isosteres, which result from the exchange of an atom or group of atoms to create a new compound with similar biological properties to the parent carboxylic acid or ester, are known in the art. The bioisosteric replacement may be physicochemically or topologically based. An example of an isosteric replacement for a carboxylic acid is CONHSO2(alkyl) such as CONHSO2Me.
It will be apparent to one skilled in the art that certain compounds of this invention may exist in tautomeric forms or hydrated forms, all such forms of the compounds being within the scope of the invention. Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by a 13Cxe2x80x94 or 14Cxe2x80x94 enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools or probes in biological assays.
One embodiment of this invention relates to compounds of formula I where the XYZ-containing ring is an isoxazole, as shown by the general formula IA below: 
where R2 is preferably alkyl, such as methyl, or CH2(heterocyclyl), such as CH2(N-morpholinyl); G is preferably Ar1; and R1 is preferably T(n)xe2x80x94Ar2 or T(n)xe2x80x94R, wherein n is most preferably zero. Most preferred are those compounds where G, R1, and R2 are as just described, and Qxe2x80x94NH is an aminopyridine or aminopyrimidine where the NH is at the 2 position of the ring: 
or Qxe2x80x94NH is an amino pyrazole: 
Table 1 below shows representative examples of IA compounds where Q is a pyrimidine, pyridine or pyrazole and R1 is Ar2, represented by formula IIA. 
For compounds of Formula IIA where R1 is phenyl, preferred phenyl substituents are selected from hydrogen and one or more halo, aliphatic, substituted aliphatic (preferably haloalkyl), alkoxy, CN, CO2H, CO2(alkyl), S(alkyl), CONH2, CO(alkyl), SO2(alkyl), CO(phenyl), or NO2. Preferred G groups are phenyl rings optionally substituted with one or more groups independently selected from alkyl, alkoxy or halogen.
Examples of compounds of Formula IIA where R1 is other than phenyl are shown below in Table 2.
Preferred IIA compounds are those where Ar1 is an unsubstituted phenyl or a phenyl substituted with one or more halo, alkyl or alkoxy. More preferred IIA compounds are those where Ar1 is as just described, and Ar2 is a naphthyl or phenyl optionally substituted with one or more halo, alkyl, alkoxy, haloalkyl, carboxyl, alkoxycarbonyl, cyano, or CONH2, or an indanone (as in compound IIAA-11). Also preferred are IIA compounds where R1 is an optionally substituted alkyl or optionally substituted cycloalkyl, more preferably alkoxyalkyl, alkoxycarbonylalkyl, hydroxyalkyl, pyridinylalkyl, alkoxycycloalkyl, alkoxycarbonylcycloalkyl, or hydroxycycloalkyl. Examples of these preferred compounds include IIAA-24, IIAA-33 through IIAA-36, IIAA-38 and IIAA-40.
One embodiment of this invention relates to compounds of formula IA where Q is a pyrimidine ring and R1 is Txe2x80x94Ar2 where T is selected from CO, CO2, CONH, S(O)2, S(O)2NH, COCH2 and CH2. When R1 is Txe2x80x94Ar2, preferred compounds are those where T is C(xe2x95x90O), represented by formula IIIA. Table 3 below shows representative examples of IIIA compounds. 
Preferred IIIA compounds are those compounds where Ar1 is an unsubstituted phenyl or a phenyl substituted with one or more substituents independently selected from halogen. More preferred IIIA compounds are those where Ar1 is just described, and Ar2 is a thienyl, an unsubstituted phenyl or a phenyl substituted with one or more substituents independently selected from halogen, alkyl, alkoxy, CO2H or CO2R.
Examples of other compounds where R1 is T-Ar1 are shown below where A is N or CH, and T is one of the following: CH2 (exemplified by IVA-1), S(O)2 (VA-1), CONH (VIA-1), COCH2 (VIIA-1), CO2, (VIIIA-1), and S(O)2NH (IXA-1). In other examples of these embodiments the phenyl rings may be optionally substituted as described above. 
Another embodiment of this invention relates to compounds of formula IA where R1 is Txe2x80x94R, R is a C3-C6 cycloalkyl ring or a C1-C6 straight chain or branched alkyl or alkenyl group optionally substituted by halogen and T is as described above. When R1 is Txe2x80x94R, preferred compounds are those where T is C(xe2x95x90O) as represented by formula XA. Table 4 below shows representative examples of XA compounds. 
Preferred R2 groups of formula I include xe2x80x94CH2OR, xe2x80x94CH2OH, xe2x80x94CH2 (heterocyclyl), xe2x80x94CH2 (substituted heterocyclyl), xe2x80x94CH2N(R)21 and an R group such as methyl. Representative examples of compounds wherein R2 is other than methyl (formula IXA) are shown in Table 5 below. 
The XYZ-containing ring of formula I may be an isoxazole ring as shown above or it may an isomeric isoxazole or xe2x80x9creversexe2x80x9d isoxazole (IB). In this embodiment Q is preferably a pyrimidine or pyridine ring where A is N or CH, or Q is a pyrazole ring, and R2 is aliphatic or substituted aliphatic. 
Examples of IB compounds are shown in Table 6 below.
In another embodiment of this invention, the XYZ-containing ring is a pyrazole ring of formula IC: 
For compounds of formula IC, G is preferably an optionally substituted aryl. Specific examples of IC compounds are shown in Table 7 below. 
Other embodiments of this invention relate to compounds where the XYZ-containing ring is a furan (ID) or a triazole (IE). These embodiments are exemplified below where R1 is phenyl, R2xe2x80x2 is hydrogen, and A is N or CH. 
For compounds of formula IB-IE, the phenyl rings of Ar1 and Ar2 may be optionally substituted as shown above for the isoxazoles of formula IA.
The compounds of this invention may be prepared in general by methods known to those skilled in the art for analogous compounds, as illustrated by the general schemes below and by the preparative examples that follow. 
Scheme I above shows a route for making isoxazoles where Q is a pyrimidine ring. The starting benzaldehyde oxime 1 may be converted to the xcex1-chlorobenzaldehyde oxime 2 using N-chlorosuccinimide and a catalytic amount of pyridine. Condensation of 2 with 2,4-pentanedione provides the isoxazole 3 which may be treated with dimethylformamide dimethylacetal to obtain the enamine 4. After an aqueous work-up and without purification, 4 may be cyclized with guanidine hydrochloride to the aminopyrimidine 5. Compounds of formula IIA may be obtained from 5 according to step (e) using the appropriate arylbromide in the presence of tris(dibenylideneacetone) dipalladium. Alternatively, 5 may be treated with the appropriate acid chloride in a pyridine/benzene solvent according to step (f) to give compounds of formula IVA. If the acid chloride is a Ar2COCl, compounds of formula IIIA may be obtained in a similar manner. 
Reagents: (a) i. LDA, ii. 2-benzyloxy-N-methoxy-N-methyl-acetamide, xe2x88x9278xc2x0 C. to rt; (b) Et3N, EtOH, rt to reflux; (c) oxone; (d) iodotrimethylsilane; (e) PPh3, CBr4; (f) morpholine, Et3N; (g) 4-aminocyclohexanol, DMSO, 80xc2x0 C.; (h) NaOEt, EtOH; (i) cyclohexylamine, DMSO, 80xc2x0 C.; (j) 3:1 trifluoroacetic acid/H2O; 100xc2x0 C.
Scheme II above shows a route for making isoxazoles of this invention where Q is a pyrimidine ring and R2 is modified by various groups. 
Scheme III above shows a synthetic route for making isoxazoles of this invention where Q is a pyridine and R2 is modified by various groups. In Scheme II and Scheme III, the isoxazole ring is first constructed and then the 2-position of the pyrimidine or pyridine ring is elaborated with the appropriate NHR1 substitution. It will be apparent to one skilled in the art that position 2 of the pyrimidine or pyridine ring can be elaborated with the appropriate NHR1 substitution before the isoxazole ring is constructed. Accordingly, isoxazoles of this invention may be obtained by performing step (b) using an appropriate intermediate having the formula XII: 
where A is N or CH; R1 and R2 are as described above; and PG is hydrogen or a nitrogen protecting group. Nitrogen protecting groups are well-known and include groups such as benzyl or CO2R, where R is preferably alkyl, allyl or benzyl. 
Scheme IV above shows a synthetic route for making reverse isoxazoles of this invention where Q is a pyrimidine ring. 
Scheme V above shows a synthetic route for making reverse isoxazoles of this invention where Q is a pyridine ring. 
Scheme VI above shows a general route for preparing compounds of this invention wherein Q is a pyrazole ring. 
Scheme VII above shows a general route for preparing compounds of this invention wherein the XYZ ring is a pyrazole ring.
Certain of the intermediates that are useful for making the kinase inhibitors of this invention are believed to be novel. Accordingly, one embodiment of this invention relates to compounds XII above and compounds represented by formula XIII: 
wherein:
Xxe2x80x94Y is Nxe2x80x94O or Oxe2x80x94N providing an isoxazole or reverse isoxazole ring;
A is N or CH;
G is R, aryl or substituted aryl;
R is aliphatic or substituted aliphatic
R2 is selected from hydrogen, xe2x80x94R, xe2x80x94CH2OR, xe2x80x94CH2OH, xe2x80x94CHxe2x95x90O, xe2x80x94CH2SR, xe2x80x94CH2S(O)2R, xe2x80x94CH2(Cxe2x95x90O)R, xe2x80x94CH2CO2R, xe2x80x94CH2CO2H, xe2x80x94CH2CN, xe2x80x94CH2NHR, xe2x80x94CH2N(R)2, xe2x80x94CHxe2x95x90Nxe2x80x94OR, xe2x80x94CHxe2x95x90NNHR, xe2x80x94CHxe2x95x90NN(R)2, xe2x80x94CHxe2x95x90NNHCOR, xe2x80x94CHxe2x95x90NNHCO2R, xe2x80x94CHxe2x95x90NNHSO2R, -aryl, -substituted aryl, xe2x80x94CH2 (aryl), xe2x80x94CH2 (substituted aryl), xe2x80x94CH2NH2, xe2x80x94CH2NHCOR, xe2x80x94CH2NHCONHR, xe2x80x94CH2NHCON(R)2, xe2x80x94CH2NRCOR, xe2x80x94CH2NHCO2R, xe2x80x94CH2CONHR, xe2x80x94CH2CON(R)2, xe2x80x94CH2SO2NH2, xe2x80x94CH2 (heterocyclyl), xe2x80x94CH2(substituted heterocyclyl), -(heterocyclyl), or -(substituted heterocyclyl); and
R1 is selected from halogen, NH2, SR, or SO2R.
The activity of the JNK inhibitors of this invention may be assayed in vitro, in vivo or in a cell line. In vitro assays include assays that determine inhibition of either the kinase activity or ATPase activity of activated JNK. For example, see the testing examples described below. Alternate in vitro assays quantitate the ability of the inhibitor to bind to JNK and may be measured either by radiolabelling the inhibitor prior to binding, isolating the inhibitor/JNK complex and determining the amount of radiolabel bound, or by running a competition experiment where new inhibitors are incubated with JNK bound to known radioligands. One may use any type or isoform of JNK, depending upon which JNK type or isoform is to be inhibited.
The JNK inhibitors or pharmaceutical salts thereof may be formulated into pharmaceutical compositions for administration to animals or humans. These pharmaceutical compositions, which comprise an amount of JNK inhibitor effective to treat or prevent a JNK-mediated condition and a pharmaceutically acceptable carrier, are another embodiment of the present invention.
The term xe2x80x9cJNK-mediated conditionxe2x80x9d, as used herein means any disease or other deleterious condition in which JNK is known to play a role. Such conditions include, without limitation, inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, cancer, infectious diseases, neurodegenerative diseases, allergies, reperfusion/ischemia in stroke, heart attacks, angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, thrombin-induced platelet aggregation, and conditions associated with prostaglandin endoperoxidase synthase-2.
Inflammatory diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, acute pancreatitis, chronic pancreatitis, asthma, allergies, and adult respiratory distress syndrome.
Autoimmune diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, glomerulonephritis, rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronic thyroiditis, Graves"" disease, autoimmune gastritis, diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopic dermatitis, chronic active hepatitis, myasthenia gravis, multiple sclerosis, inflammatory bowel disease, ulcerative colitis, Crohn""s disease, psoriasis, or graft vs. host disease.
Destructive bone disorders which may be treated or prevented by the compounds of this invention include, but are not limited to, osteoporosis, osteoarthritis and multiple myeloma-related bone disorder.
Proliferative diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, acute myelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma, Kaposi""s sarcoma, multiple myeloma and HTLV-1 mediated tumorigenesis.
Angiogenic disorders which may be treated or prevented by the compounds of this invention include solid tumors, ocular neovasculization, infantile haemangiomas. Infectious diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, sepsis, septic shock, and Shigellosis.
Viral diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, acute hepatitis infection (including hepatitis A, hepatitis B and hepatitis C), HIV infection and CMV retinitis.
Neurodegenerative diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, Alzheimer""s disease, Parkinson""s disease, amyotrophic lateral sclerosis (ALS), epilepsy, seizures, Huntington""s disease, traumatic brain injury, ischemic and hemorrhaging stroke, cerebral ischemias or neurodegenerative disease, including apoptosis-driven neurodegenerative disease, caused by traumatic injury, acute hypoxia, ischemia or glutamate neurotoxicity.
xe2x80x9cJNK-mediated conditionsxe2x80x9d also include ischemia/reperfusion in stroke, heart attacks, myocardial ischemia, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, hepatic ischemia, liver disease, congestive heart failure, pathologic immune responses such as that caused by T cell activation and thrombin-induced platelet aggregation.
In addition, JNK inhibitors of the instant invention may be capable of inhibiting the expression of inducible pro-inflammatory proteins. Therefore, other xe2x80x9cJNK-mediated conditionsxe2x80x9d which may be treated by the compounds of this invention include edema, analgesia, fever and pain, such as neuromuscular pain, headache, cancer pain, dental pain and arthritis pain.
The compounds of this invention are also useful as inhibitors of Src-family kinases, especially Src and Lck. For a general review of these kinases see Thomas and Brugge, Annu. Rev. Cell Dev. Biol. (1997) 13, 513; Lawrence and Niu, Pharmacol. Ther. (1998) 77, 81; Tatosyan and Mizenina, Biochemistry (Moscow) (2000) 65, 49. Accordingly, these compounds are useful for treating diseases or conditions that are known to be affected by the activity of one or more Src-family kinases. Such diseases or conditions include hypercalcemia, restenosis, hypercalcemia, osteoporosis, osteoarthritis, symptomatic treatment of bone metastasis, rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, psoriasis, lupus, graft vs. host disease, T-cell mediated hypersensitivity disease, Hashimoto""s thyroiditis, Guillain-Barre syndrome, chronic obtructive pulmonary disorder, contact dermatitis, cancer, Paget""s disease, asthma, ischemic or reperfusion injury, allergic disease, atopic dermatitis, and allergic rhinitis. Diseases that are affected by Src activity, in particular, include hypercalcemia, osteoporosis, osteoarthritis, cancer, symptomatic treatment of bone metastasis, and Paget""s disease. Diseases that are affected by Lck activity, in particular, include autoimmune diseases, allergies, rheumatoid arthritis, and leukemia. Compounds of formula II-A and I-B wherein Ar2 is aryl are especially useful for treating diseases associated with the Src-family kinases, particularly Src or Lck.
In addition to the compounds of this invention, pharmaceutically acceptable derivatives or prodrugs of the compounds of this invention may also be employed in compositions to treat or prevent the above-identified disorders.
A xe2x80x9cpharmaceutically acceptable derivative or prodrugxe2x80x9d means any pharmaceutically acceptable salt, ester, salt of an ester or other derivative of a compound of this invention which, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof. Particularly favored derivatives or prodrugs are those that increase the bioavailability of the compounds of this invention when such compounds are administered to a mammal (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment (e.g., the brain or lymphatic system) relative to the parent species.
Pharmaceutically acceptable prodrugs of the compounds of this invention include, without limitation, esters, amino acid esters, phosphate esters, metal salts and sulfonate esters.
Pharmaceutically acceptable salts of the compounds of this invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.
Salts derived from appropriate bases include alkali metal (e.g., sodium and potassium), alkaline earth metal (e.g., magnesium), ammonium and N+(C1-4 alkyl)4 salts. This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization.
Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.
The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term xe2x80x9cparenteralxe2x80x9d as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously.
Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer""s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.
The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
Alternatively, the pharmaceutical compositions of this invention may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.
The pharmaceutical compositions of this invention may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.
Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.
For topical applications, the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petrolatum.
The pharmaceutical compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
The amount of JNK inhibitor that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular mode of administration. Preferably, the compositions should be formulated so that a dosage of between 0.01-100 mg/kg body weight/day of the inhibitor can be administered to a patient receiving these compositions.
It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of inhibitor will also depend upon the particular compound in the composition.
According to another embodiment, the invention provides methods for treating or preventing a JNK-mediated condition comprising the step of administering to a patient one of the above-described pharmaceutical compositions. The term xe2x80x9cpatientxe2x80x9d, as used herein, means an animal, preferably a human.
Preferably, that method is used to treat or prevent a condition selected from inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, infectious diseases, degenerative diseases, neurodegenerative diseases, allergies, reperfusion/ischemia in stroke, heart attacks, angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiac hypertrophy, and thrombin-induced platelet aggregation, or any specific disease or disorder described above.
Depending upon the particular JNK-mediated condition to be treated or prevented, additional drugs, which are normally administered to treat or prevent that condition, may be administered together with the inhibitors of this invention. For example, chemotherapeutic agents or other anti-proliferative agents may be combined with the JNK inhibitors of this invention to treat proliferative diseases.
Those additional agents may be administered separately, as part of a multiple dosage regimen, from the JNK inhibitor-containing composition. Alternatively, those agents may be part of a single dosage form, mixed together with the JNK inhibitor in a single composition.
In order that the invention described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner.